We have shown that cluster intradermal vaccination of CRT/E7DNA (with a short interval between vaccinations) more rapidly induced E7-specific immune responses and generated better therapeutic effects against E7-expresslng tumors compared to DNA vaccination with longer intervals. These findings prompted us to propose a phase I trial to investigate whether the repeated, cluster intradermal CRT/E7 DNA vaccination is safe and can generate E7-specific CD8+ T cell immune responses in patients with HPV-16 associated stage 1B1 resectable cervical cancer. We plan to vaccinate two cohorts of patients, one before and one after tumor resection. The proposed trial using cluster vaccination regimen permits us to complete the vaccination regimen before tumor resection allowing us to assess the influence of DNA vaccination on tumor microenvironment without compromising the standard care. Clinical grade pNGVL4a-CRT/E7(detox) DNA vaccine has been manufactured by NCI RAID program and formulated in the proprietary ND-10 gene gun device by PowderMed/Pfizer for intradermal vaccination. The proposed trial will be performed at the University of Alabama at Birmingham, which has one of the highest populations of stage IBl cervical cancer patients. Specifically, we plan to: Aim 1 Evaluate the safety/toxiclty associated with repeated, cluster intradermal pNGVL4a-CRT/E7 (detox) DNA vaccination via gene gun in patients with HPV 16-associated stage IBl cervical cancer, using a dose-escalating regimen;Aim 2. Characterize systemic HPV-16 E6 and E7-speciflc humoral and T cell-mediated immune responses in stage IBl cervical cancer patients receiving the DNA vaccination via gene gun;Aim 3. Characterize the presence of HPV-16 E6 and E7-speclfic CD8+ T cell immune responses in the tumor and draining lymph nodes in selected stage IBl cervical cancer patients receiving the DNA vaccination via gene gun;Aim 4. Determine the subset population of immune cells infiltrating the tumor bed, B7H-1 and STAT-3 expression in tumor microenvironment and to determine the apoptotic tumor cell death in stage IB1 cervical cancer patients receiving the DNA vaccination via gene gun. The successful implementation of the proposed study will allow us to assess the systemic and local antigen- specific immune responses following repeated cluster intradermal DNA vaccination using the ND-10 gene gun device in HPV-16 associated stage IBl cervical cancer. RELEVANCE (See instructions): This project aims to develop a therapeutic HPV DNA vaccine formulated in a gene gun device for the treatment of patients with HPV-16 associated stage IBl cervical cancer.